The cytoskeleton dynamics-dependent LINC complex in periodontal ligament stem cells transmits mechanical stress to the nuclear envelope and promotes YAP nuclear translocation.

BACKGROUND: Periodontal ligament stem cells (PDLSCs) are important seed cells in tissue engineering and clinical applications. They are the priority receptor cells for sensing various mechanical stresses. Yes-associated protein (YAP) is a recognized mechanically sensitive transcription factor. However, the role of YAP in regulating the fate of PDLSCs under tension stress (TS) and its underlying mechanism is still unclear. METHODS: The effects of TS on the morphology and fate of PDLSCs were investigated using fluorescence staining, transmission electron microscopy, flow cytometry and quantitative real-time polymerase chain reaction (qRT-PCR). Then qRT-PCR, western blotting, immunofluorescence staining and gene knockdown experiments were performed to investigate the expression and distribution of YAP and its correlation with PDLSCs proliferation. The effects of cytoskeleton dynamics on YAP nuclear translocation were subsequently explored by adding cytoskeleton inhibitors. The effect of cytoskeleton dynamics on the expression of the LINC complex was proved through qRT-PCR and western blotting. After destroying the LINC complex by adenovirus, the effects of the LINC complex on YAP nuclear translocation and PDLSCs proliferation were investigated. Mitochondria-related detections were then performed to explore the role of mitochondria in YAP nuclear translocation. Finally, the in vitro results were verified by constructing orthodontic tooth movement models in Sprague-Dawley rats. RESULTS: TS enhanced the polymerization and stretching of F-actin, which upregulated the expression of the LINC complex. This further strengthened the pull on the nuclear envelope, enlarged the nuclear pore, and facilitated YAP's nuclear entry, thus enhancing the expression of proliferation-related genes. In this process, mitochondria were transported to the periphery of the nucleus along the reconstructed microtubules. They generated ATP to aid YAP's nuclear translocation and drove F-actin polymerization to a certain degree. When the LINC complex was destroyed, the nuclear translocation of YAP was inhibited, which limited PDLSCs proliferation, impeded periodontal tissue remodeling, and hindered tooth movement. CONCLUSIONS: Our study confirmed that appropriate TS could promote PDLSCs proliferation and periodontal tissue remodeling through the mechanically driven F-actin/LINC complex/YAP axis, which could provide theoretical guidance for seed cell expansion and for promoting healthy and effective tooth movement in clinical practice.

Effects of different designs of orthodontic clear aligners on the maxillary central incisors in the tooth extraction cases: a biomechanical study.

BACKGROUND: Controlling the 3D movement of central incisors during tooth extraction cases with clear aligners is important but challenging in invisible orthodontic treatment. This study aimed to explore the biomechanical effects of central incisors in tooth extraction cases with clear aligners under different power ridge design schemes and propose appropriate advice for orthodontic clinic. METHODS: A series of Finite Element models was constructed to simulate anterior teeth retraction or no retraction with different power ridge designs. These models all consisted of maxillary dentition with extracted first premolars, alveolar bone, periodontal ligaments and clear aligner. And the biomechanical effects were analysed and compared in each model. RESULTS: For the model of anterior teeth retraction without power ridge and for the model of anterior teeth no retraction with a single power ridge, the central incisors exhibited crown lingual inclination and relative extrusion. For the model of anterior teeth no retraction with double power ridges, the central incisors tended to have crown labial inclination and relative intrusion. For the model of anterior tooth retraction with double power ridges, the central incisors exhibited a similar trend to the first kind of model, but as the depth of the power ridge increased, there was a gradual decrease in crown retraction value and an increase in crown extrusion value. The simulated results showed that von-Mises stress concentration was observed in the cervical and apical regions of the periodontal ligaments of the central incisors. The clear aligner connection areas of adjacent teeth and power ridge areas also exhibited von-Mises stress concentration and the addition of power ridge caused the clear aligner to spread out on the labial and lingual sides. CONCLUSIONS: The central incisors are prone to losing torque and extruding in tooth extraction cases. Double power ridges have a certain root torque effect when there are no auxiliary designs, but they still cannot rescue tooth inclination during tooth retraction period. For tooth translation, it may be a better clinical procedure to change the one-step aligner design to two-step process: tilting retraction and root control.

MiR-134-5p/Stat3 Axis Modulates Proliferation and Migration of MSCs Co-Cultured with Glioma C6 Cells by Regulating Pvt1 Expression.

Mesenchymal stem cells (MSCs) are critical in regenerating tissues because they can differentiate into various tissue cells. MSCs interact closely with cells in the tissue microenvironment during the repair of damaged tissue. Although regarded as non-healing wounds, tumors can be treated by MSCs, which showed satisfactory treatment outcomes in previous reports. However, it is largely unknown whether the biological behaviors of MSCs would be affected by the tumor microenvironment. Exploring the truth of tumor microenvironmental cues driving MSCs tumor "wound" regeneration would provide a deeper understanding of the biological behavior of MSCs. Therefore, we mimicked the tumor microenvironment using co-cultured glioma C6 cells and rat MSCs, aiming to assess the proliferation and migration of MSCs and the associated effects of Stat3 in this process. The results showed that co-cultured MSCs significantly exhibited enhanced tumorigenic, migratory, and proliferative abilities. Both up-regulation of Stat3 and down-regulation of miR-134-5p were detected in co-cultured MSCs. Furthermore, miR-134-5p directly regulated Stat3 by binding to the sequence complementary to microRNA response elements in the 3'-UTR of its mRNA. Functional studies showed that both the migration and proliferation abilities of co-cultured MSCs were inhibited by miR-134-5p, whereas Stat3 gain-of-function treatment reversed these effects. In addition, Pvt1 was confirmed to be regulated by miR-134-5p through Stat3 and the suppression of Pvt1 reduced the migration and proliferation abilities of co-cultured MSCs. To sum up, these results demonstrate a suppressive role of miR-134-5p in tumor-environment-driven malignant transformation of rat MSCs through directly targeting Stat3, highlighting a crucial role of loss-of-function of miR-134-5p/Stat3 axis in the malignant transformation, providing a reference to the potential clinic use of MSCs.

Improved lip esthetics in patients with skeletal Class III malocclusion and facial asymmetry after isolated mandibular orthognathic surgery.

INTRODUCTION: Asymmetry of the lips severely affects facial esthetics and is often one of the chief complaints of orthognathic patients, especially those with Class III malocclusion. The objectives of this study were to investigate the changes in lip symmetry in patients with mandibular prognathism and deviation and the relationships between jaw hard tissue and lip soft-tissue changes. METHODS: Three-dimensional facial scan and cone-beam computed tomography scan data of 30 orthodontic-orthognathic patients treated with bilateral sagittal split ramus osteotomy were combined to conduct the research. Paired-sample t test and Pearson correlation coefficient were applied to compare the differences in the same variable before and after the orthognathic surgery and the potential correlations between the changes in hard and soft variables. To explore the important hard tissue variables influencing the lip soft-tissue changes, linear regression analysis was performed. RESULTS: Although there was significant upper lip asymmetry presurgery, the upper lip asymmetry was corrected postsurgery. Surgical correction of the mandibular deviation was also accompanied by lengthening of the bilateral philtrum crests. Improvement in lip asymmetry and lengthening of the philtrum crests were primarily related to the transverse correction of the mandible rather than sagittal changes. The corresponding prediction formulas were established. CONCLUSIONS: The isolated mandibular bilateral sagittal split ramus osteotomy surgery can substantially improve the upper and lower lip asymmetry in patients with mandibular prognathism and deviation, but one should be wary of the unesthetic effects associated with lengthening of the philtrum crests.

Reliability of cone beam CT for morphometry of nasolabial soft tissue in patients with skeletal class III malocclusion: A qualitative and quantitative analysis.

OBJECTIVE: To assess reliability of cone-beam CT (CBCT) for nasolabial soft tissue measurements in patients with skeletal class III malocclusion based on 3-dimensional (3D) facial scanner results. METHODS: CBCT and 3D facial scan images of 20 orthognathic patients are used in this study. Eleven soft tissue landmarks and 15 linear and angular measurements are identified and performed. For qualitative evaluation, Shapiro-Wilk test and Bland-Altman plots are applied to analyze the equivalence of the measurements derived from these two kinds of images. To quantify specific deviation of CBCT measurements from facial scanner, the latter is set as a benchmark, and mean absolute difference (MAD) and relative error magnitude (REM) for each variable are also calculated. RESULTS: Statistically significant differences are observed in regions of nasal base and lower lip vermilion between two methods. MAD value for all length measurements are less than 2 mm and for angular variables < 8°. The average MAD and REM for length measurements are 0.94 mm and 5.64%, and for angular measurements are 2.27° and 3.78%, respectively. CONCLUSIONS: The soft tissue results measured by CBCT show relatively good reliability and can be used for 3D measurement of soft tissue in the nasolabial region clinically.

Three-dimensional analysis of lip soft tissue changes and related jaw changes in patients with skeletal class III malocclusion and facial asymmetry.

OBJECTIVE: To investigate three-dimensional soft tissue changes of lips and related jaw changes in patients with skeletal class III malocclusion and facial asymmetry using cone-beam computerized tomography (CBCT). METHOD: This study included 46 CBCT scans of patients with aforementioned character before (T1) and 6-12 months after orthognathic surgery (T2). Subjects were divided into 2 groups according to two types of orthognathic surgery namely, the one-jaw surgery group who underwent bilateral sagittal splint ramus osteotomy (BSSRO) and the two-jaw surgery group who underwent Le Fort I osteotomy and BSSRO. Mimics 19.0 software are used for model reconstruction, landmark location and three-dimensional cephalometric analysis. Descriptive statistics and correlation analyses are used to investigate jaw hard tissue and lip soft tissue changes. RESULTS: In one-jaw group, the mandible shows changes in contour and position (p < 0.05), and the surgery causes changes of lip structure on the deviated side. While in two-jaw group, jaws only show changes in spatial position, and surgery changes contour of bilateral lips and nasolabial angle (p < 0.05). At the same time, lip symmetry increases significantly in both groups postoperatively. CONCLUSIONS: Orthognathic surgery can improve lip aesthetics in patients with skeletal class III malocclusion and facial asymmetry. However, changes induced by two surgical approaches are different. Surgeons should have a clear acquaintance with this difference to deal with different situations.

[PVT1 Promoted the Proliferation and Migration Ability of BMSCs in Glioma C6 Microenvironment].

OBJECTIVE: To investigate the changes in the proliferation and migration ability of bone marrow mesenchymal stem cells (BMSCs) after indirect co-culturing with glioma C6 cells, and to examine the role of plasmacytoma variant translocation 1 gene ( PVT1), a long non-coding RNA (lncRNA), in these changes. METHODS: After separation, cultivation and identification of BMSCs, BMSCs of good growth condition were picked out and indirectly co-cultured with glioma C6 cells in Transwell chambers. These cells are henceforth referred to as the co-culture group. Normal BMSCs cultured separately were the control group. CCK-8 and soft agar colony formation assay were used to examine the proliferation ability of the two groups of cells. Flow cytometry was used to examine the cell cycle. Wound healing assay and Transwell assay were used to explore the migration ability of the cells. Quantitative real-time PCR (qRT-PCR) was used to examine the genetic expression level of PVT1 in the two groups. The above-mentioned tests were repeated after the co-cultured BMSCs were transfected with si- PVT1 (si- PVT1 group) and si-NC (si-NC group). In addition, qRT-PCR was done to evaluate the expression of CyclinD1, a cell cycle protein gene, and matrix metalloproteinases 2 and 9 ( MMP2 and MMP9), the migration-related genes in the si- PVT1 and si-NC transfected co-cultured BMSCs. RESULTS: The BMSCs used in the present study possess the capability of osteogeneic and adipogenic differentiation. Compared with the control group, the co-cultured BMSCs had smaller size, disorderly arrangement and the lack of intercellular contact inhibition. The proliferation and migration ability was significantly enhanced, the proportions of S and G 2 phase cells greatly increased and the expression level of PVT1 was significantly up-regulated ( P<0.05) in the co-cultured group in comparison with those of the control group. When compared with the si-NC group, the si- PVT1 group showed inhibited proliferation and migration ability of the co-cultured BMSCs; the percentage of G 1 phase cells increased, while that of S phase decreased; the expression of PVT1, CyclinD1, MMP2 and MMP9 mRNA also decreased ( P<0.05) in the si- PVT1 group. CONCLUSION: The enhanced proliferation and migration ability of BMSCs in the glioma C6 microenvironment may be associated with the up-regulated expression of PVT1 .